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A mechanistic study on SMOB-ADP1: an NADH:flavin oxidoreductase of the two-component styrene monooxygenase of Acinetobacter baylyi ADP1

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Abstract

Two styrene monooxygenase types, StyA/StyB and StyA1/StyA2B, have been described each consisting of an epoxidase and a reductase. A gene fusion which led to the chimeric reductase StyA2B and the occurrence in different phyla are major differences. Identification of SMOA/SMOB-ADP1 of Acinetobacter baylyi ADP1 may enlighten the gene fusion event since phylogenetic analysis indicated both proteins to be more related to StyA2B than to StyA/StyB. SMOB-ADP1 is classified like StyB and StyA2B as HpaC-like reductase. Substrate affinity and turnover number of the homo-dimer SMOB-ADP1 were determined for NADH (24 µM, 64 s−1) and FAD (4.4 µM, 56 s−1). SMOB-ADP1 catalysis follows a random sequential mechanism, and FAD fluorescence is quenched upon binding to SMOB-ADP1 (K d = 1.8 µM), which clearly distinguishes that reductase from StyB of Pseudomonas. In summary, this study confirmes made assumptions and provides phylogenetic and biochemical data for the differentiation of styrene monooxygenase-related flavin reductases.

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Acknowledgments

The project was financed by the Saxon Ministry for Environment, Agriculture, and Geology (LfULG-1771508003). Dirk Tischler was supported by the European Social Fund and the Saxon Government (GETGEOWEB: 100101363). We are grateful to Adrie Westphal for advice during fluorescence measurements.

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  1. Environmental Microbiology Group, Interdisciplinary Ecological Center, TU Bergakademie Freiberg, Leipziger Str. 29, 09599, Freiberg, Germany

    Janosch A. D. Gröning, Stefan R. Kaschabek, Michael Schlömann & Dirk Tischler

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  1. Janosch A. D. Gröning
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Correspondence to Dirk Tischler.

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Communicated by Erko Stackebrandt.

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Gröning, J.A.D., Kaschabek, S.R., Schlömann, M. et al. A mechanistic study on SMOB-ADP1: an NADH:flavin oxidoreductase of the two-component styrene monooxygenase of Acinetobacter baylyi ADP1. Arch Microbiol 196, 829–845 (2014). https://doi.org/10.1007/s00203-014-1022-y

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